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RNA sequences upstream of the 3' splice site repress splicing of mutantyeast ACT1 introns.
RNA. 1996 May; 2(5):492-505.RNA

Abstract

A yeast ACT1 intron in which both the first and last intron nucleotides are mutated, the /a-c/ intron, splices 10% as well as wild type. We selected for additional cis-acting mutations that improve the splicing of /a-c/ introns and recovered small deletions upstream of the 3' splice site. For example, deletion of nucleotides -9 and -10 upstream of the 3' splice site increased the splicing activity of the /a-c/ intron to 30% that of the wild-type ACT1 intron. To determine if the increased /a-c/ splicing was due to changes in intron spacing or sequence, we made mutations that mimicked the local sequence of the delta-9, -10 deletion without deleting any nucleotides. These mutants also increased /a-c/ splicing, indicating that the increased splicing activity was due to changes in intron sequence. The delta-9, -10 deletion was not allele specific to the /a-c/ intron, and improved the splicing efficiency of many mutant introns with step II splicing defects. To further define the sequences required for improved splicing of mutant introns, we randomized the region upstream of the ACT1 3' splice site. We found that almost all sequence alterations improved the splicing of the /a-c/ intron. We postulate that this sequence near the 3' end of the intron represses the splicing of mutant introns, perhaps by serving as the binding site for a negative splicing factor.

Authors+Show Affiliations

Department of Biochemistry, University of Minnesota, Minneapolis, 55455, USA.No affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

8665416

Citation

Kivens, W, and P G. Siliciano. "RNA Sequences Upstream of the 3' Splice Site Repress Splicing of Mutantyeast ACT1 Introns." RNA (New York, N.Y.), vol. 2, no. 5, 1996, pp. 492-505.
Kivens W, Siliciano PG. RNA sequences upstream of the 3' splice site repress splicing of mutantyeast ACT1 introns. RNA. 1996;2(5):492-505.
Kivens, W., & Siliciano, P. G. (1996). RNA sequences upstream of the 3' splice site repress splicing of mutantyeast ACT1 introns. RNA (New York, N.Y.), 2(5), 492-505.
Kivens W, Siliciano PG. RNA Sequences Upstream of the 3' Splice Site Repress Splicing of Mutantyeast ACT1 Introns. RNA. 1996;2(5):492-505. PubMed PMID: 8665416.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - RNA sequences upstream of the 3' splice site repress splicing of mutantyeast ACT1 introns. AU - Kivens,W, AU - Siliciano,P G, PY - 1996/5/1/pubmed PY - 1996/5/1/medline PY - 1996/5/1/entrez SP - 492 EP - 505 JF - RNA (New York, N.Y.) JO - RNA VL - 2 IS - 5 N2 - A yeast ACT1 intron in which both the first and last intron nucleotides are mutated, the /a-c/ intron, splices 10% as well as wild type. We selected for additional cis-acting mutations that improve the splicing of /a-c/ introns and recovered small deletions upstream of the 3' splice site. For example, deletion of nucleotides -9 and -10 upstream of the 3' splice site increased the splicing activity of the /a-c/ intron to 30% that of the wild-type ACT1 intron. To determine if the increased /a-c/ splicing was due to changes in intron spacing or sequence, we made mutations that mimicked the local sequence of the delta-9, -10 deletion without deleting any nucleotides. These mutants also increased /a-c/ splicing, indicating that the increased splicing activity was due to changes in intron sequence. The delta-9, -10 deletion was not allele specific to the /a-c/ intron, and improved the splicing efficiency of many mutant introns with step II splicing defects. To further define the sequences required for improved splicing of mutant introns, we randomized the region upstream of the ACT1 3' splice site. We found that almost all sequence alterations improved the splicing of the /a-c/ intron. We postulate that this sequence near the 3' end of the intron represses the splicing of mutant introns, perhaps by serving as the binding site for a negative splicing factor. SN - 1355-8382 UR - https://www.unboundmedicine.com/medline/citation/8665416/RNA_sequences_upstream_of_the_3'_splice_site_repress_splicing_of_mutantyeast_ACT1_introns_ L2 - http://www.rnajournal.org/cgi/pmidlookup?view=long&pmid=8665416 DB - PRIME DP - Unbound Medicine ER -